Cutting edge Seminar
Speaker: Ryuji Morizane (Associate Biologist, Renal Division,Brigham and Women`s Hospital
Instructor, Department of medicine, Harvard Medical School)
Date&Time: 15 Jun. (Wed.) 2016, 12:00- 13:00
Venue: Conference Room(1F), IMEG
Abstract:
Significant advances have been made within the past decade that draw upon our knowledge of kidney development to differentiate human pluripotent stem cells (hPSCs) into cells of the kidney lineage. By recapitulating metanephric kidney development in vitro, we generated nephron progenitor cells (NPCs) with 90% purity within 9 days of differentiation without additional subpopulation selection during the directed differentiation. hPSC-derived NPCs possess the developmental potential of their in vivo counterparts, forming renal vesicles that self-pattern into nephron structures. In both 2D and 3D culture, NPCs form nephron organoids containing epithelial nephron-like structures expressing markers of podocytes, proximal tubules, loops of Henle, and distal tubules in an organized, continuous arrangement that resembles the nephron in vivo.
Recent advances in genome editing using CRISPR/Cas9 systems provide the tools to generate specific mutations at desired sites. Genetics is a dominant factor contributing to variation in the differentiation and functional characteristics of differentiated hPSCs. The CRISPR/Cas9 system enables the generation of specific mutations in target genes in normal hPSCs. Kidney organoids differentiated from these cells can then be used to model kidney disease known to be caused by these genetic modifications. The parental hPSC line will be an ideal control for these studies since all genes, except for those targeted for modification, will be the same. We reported the feasibility of modeling autosomal dominant polycystic kidney disease (ADPKD) using normal hPSCs and CRISPR/Cas9 genome editing.
These novel technologies contribute to developing novel platforms for studies of human kidney development, modeling of kidney diseases, nephrotoxicity of drugs, and kidney regeneration, and provide a system in vitro for the study of intracellular and kidney compartmental interactions using differentiated human cells in an appropriate nephron context.
References:
1) Morizane R, Lam AQ, Freedman BS, Kishi S, Valerius MT, Bonventre JV. Nephron organoids derived from human pluripotent stem cells model kidney development and injury. Nat Biotechnol. 2015 Nov;33(11):1193-200.
2) Freedman BS, Brooks CR, Lam AQ, Fu H, Morizane R et al. Modelling kidney disease with CRISPR-mutant kidney organoids derived from human pluripotent epiblast spheroids. Nat Commun. 2015 Oct 23;6:8715. doi: 10.1038/ncomms9715.
3) Morizane R, Lam AQ, Freedman BS (co-first authors), Lerou PH, Valerius MT, Bonventre JV. Rapid and efficient differentiation of human pluripotent stem cells into intermediate mesoderm that forms tubules expressing kidney proximal tubular markers. J Am Soc Nephrol. 2014 Jun;25(6):1211-25.
4) Morizane R, Monkawa T, Fujii S, Yamaguchi S, Homma K, Matsuzaki Y, Okano H, Itoh H. Kidney specific protein-positive cells derived from embryonic stem cells reproduce tubular structures in vitro and differentiate into renal tubular cells. PLoS One. 2013 Jun 3;8(6):e64843.
5) Morizane R, Monkawa T, Itoh H. Differentiation of murine embryonic stem and induced pluripotent stem cells to renal lineage in vitro. Biochem Biophys Res Commun. 2009 Dec 25;390(4):1334-9.
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